Cell movement device and method

Abstract

This cell movement device moves a plurality of cells accommodated in a first container to a second container that retains a liquid. The cell movement device comprises: a head to which a tip having a leading end for sucking and discharging the cells is mounted, and which performs an operation of sucking the plurality of cells by means of the tip in the first container, moving to the second container, and discharging the plurality of cells in the tip into the second container; and a control unit that controls the operations of the head. The control unit controls the head so as to discharge the cells from the tip while moving the head in a state in which the leading end of the tip is immersed in the liquid in the second container.

Description

Cell migration device and method

[0001] The present invention relates to a cell transfer device and a cell transfer method for transferring cells contained in a first container to a second container.

[0002] For example, in the fields of medical and biological research, there is a need to move cells, such as single cells or cell colonies, from a culture vessel in which the cells are cultured to a work vessel where the cells are examined, observed, etc. For this purpose, a cell moving device is known that uses a head equipped with a suction tip to pick and hold target cells from the culture vessel and then release the held cells into the work vessel (see, for example, Patent Document 1).

[0003] When the number of target cells is very small relative to the total number of cells contained in a culture vessel, it is difficult to selectively pick the target cells. In this case, the limiting dilution method is applied to reduce the number of cells, i.e., to dilute the cell concentration, before picking the target cells. When the total number of cells in a culture vessel is large, the limiting dilution process requires a great deal of effort.

[0004] Patent No. 6181871

[0005] An object of the present invention is to provide a cell migration device and method that can efficiently pick a target cell from among a large number of cells.

[0006] A cell movement device according to one aspect of the present invention is a cell movement device that moves a plurality of cells contained in a first container to a second container that stores a liquid, and is equipped with a head that is equipped with a tip having a tip that aspirates and ejects cells, and that aspirates a plurality of cells in the first container into the tip, moves them to the second container, and ejects the plurality of cells in the tip into the second container, and a control unit that controls the operation of the head, and the control unit controls the head to eject cells from the tip while moving the head with the tip of the tip immersed in the liquid in the second container.

[0007] Another aspect of the present invention is a cell transfer method for transferring a plurality of cells contained in a first container to a second container that stores a liquid, in which a tip having a tip for aspirating and discharging cells is used to aspirate a plurality of cells into the tip in the first container, the tip is moved to the second container, and the cells are discharged from the tip while the tip is moved with the tip immersed in the liquid in the second container.

[0008] FIG. 1 is a schematic diagram illustrating an example of the configuration of a cell migration device according to an embodiment of the present invention. FIG. 2(A) is a top view of a 6-well plate, and FIG. 2(B) is a diagram illustrating the grid on the bottom of each well. FIG. 3(A) is a cross-sectional view of a tip attached to a head, and a diagram illustrating the tip's migration mechanism and suction mechanism. FIG. 3(B) is a cross-sectional view of the tip during suction operation. FIG. 3(C) is an enlarged view of the main part of FIG. 3(B). FIG. 4 is a block diagram illustrating the electrical configuration of the cell migration device. FIG. 5 is a cross-sectional view illustrating the state of cell ejection from the tip to a well. FIGS. 6(A) to 6(E) are schematic diagrams illustrating an example of tip movement during cell ejection. FIGS. 7(A) to 7(D) are cross-sectional views illustrating the process from seeding cells into a first container to picking target cells from a second container. FIGS. 8(A) to 8(D) are diagrams illustrating an example of seeding cells into a first container. FIGS. 9(A) to 9(C) are diagrams illustrating an example of cell ejection that prevents the formation of cell-undispersed regions in the second container. FIG. 10 is a flowchart showing an example of a cell picking process.

[0009] Embodiments of the present invention are described in detail below with reference to the accompanying drawings. The cell migration device and method according to the present invention can pick and move various types of cells derived from living organisms. Examples of cells derived from living organisms include single cells such as blood cells, single cells, and fertilized eggs, circulating tumor cells, small tissue fragments such as histoculture, cell aggregates such as spheroids and organoids, individual cells such as zebrafish and nematodes, and 2D or 3D cell colonies. The term "cell" used herein encompasses these various types of cells. The cell migration device of the present invention is particularly suitable for picking and moving single cells, cell aggregates, cell colonies, and other cells that generally require picking under a microscope.

[0010] [Overall Configuration of Cell Migration Apparatus] Figure 1 is a diagram showing the overall configuration of a cell migration apparatus S according to an embodiment of the present invention. Here, a cell migration apparatus S is shown that moves cells C between two containers, i.e., between a sorting plate 2 and a destination plate 4 (third container). Of course, the cell migration apparatus S may also be configured to move cells C between three or more containers.

[0011] The cell movement device S includes a light-transmitting base 1 having a horizontal mounting surface, a camera unit 5 (camera) disposed below the base 1, and a head unit 6 disposed above the base 1. A sorting plate 2, which is the source of the cells C, is mounted at a first mounting position P1 on the base 1, and a destination plate 4, which is the destination of the cells C, is mounted at a second mounting position P2. The head unit 6 includes multiple heads 61 that can move up and down in the Z direction (up and down). These heads 61 can operate simultaneously or individually. A chip 10 that aspirates and discharges the cells C is attached to the bottom end of each head 61. The camera unit 5 and head unit 6 can move in the X direction (horizontal direction) and the direction perpendicular to the plane of FIG. 1 (Y direction). The sorting plate 2 and destination plate 4 are mounted on the top surface of the base 1 within the movable range of the head unit 6.

[0012] Generally speaking, the cell movement device S performs picking by individually aspirating specific target cells C into each of a plurality of tips 10 from a sorting plate 2 on which a large number of cells C are cultured. In this embodiment, cell movement is also performed between a plurality of wells 3 provided in the sorting plate 2 to isolate the target cells C. The picked cells C are then moved to a destination plate 4, and the cells C are discharged from the plurality of tips 10 into the destination plate 4 (wells 41). Before picking the cells C, the camera unit 5 captures images of the cells C held on the sorting plate 2, and a sorting operation is performed to select good quality target cells C to be moved to the destination plate 4.

[0013] Each part of the cell migration device S will be described below. The base 1 is a flat plate having a predetermined rigidity and formed in part or in whole from a light-transmitting material. A preferred base 1 is a glass plate. By forming the base 1 from a light-transmitting material such as a glass plate, it becomes possible for the camera unit 5, located below the base 1, to capture images of the sorting plate 2 and destination plate 4, located on the upper surface of the base 1, through the base 1.

[0014] The sorting plate 2 is a container from which the cells C are moved and includes a plurality of wells 3 for culturing the cells C. Each well 3 is a small container with an open top and a flat bottom 31. A liquid medium LCM is poured into the well 3, and test cells are seeded therein. The seeded cells C are arranged on the bottom 31 in an adherent or floating state. The sorting plate 2 is made of a member made of a light-transmitting resin material or glass, so that the cells C can be imaged by a camera unit 5 disposed below. The sorting plate 2 may be, for example, a commercially available 6-well plate (e.g., Corning Model No. 3516).

[0015] FIG. 2(A) is a plan view showing an example of a sorting plate 2 consisting of the above-mentioned 6-well plate, and FIG. 2(B) is a plan view showing the bottom surface 31 of each well 3. The sorting plate 2 has six wells 3 arranged in two rows and three columns. The wells 3 are cylindrical recesses with an open top. These six wells 3 are given identification codes A1, A2, A3, B1, B2, and B3 as addresses. The bottom surface 31 of each well 3 is provided with a large number of grids 3G (multiple recesses), which are tiny recesses. The grids 3G are rectangular parallelepipeds with sides of approximately 200 μm and a height of approximately 100 μm, and are arranged in a matrix across the entire bottom surface 31. Cells C are accommodated in the grids 3G.

[0016] In this embodiment, to isolate target cells C, cells C are seeded in the first well 3A1 (first container) of the sorting plate 2, and a cell group containing the target cells C is aspirated from the grid 3G of the first well 3A1 using the chip 10 and discharged into the second well 3B1 (second container) that stores a liquid culture medium. This discharge causes the target cells C to be individually contained in one of the grids 3G of the second well 3B1. The target cells C isolated on the grid 3G are then identified. The target cells C are aspirated from the grid 3G using the chip 10 and discharged into the destination plate 4. This operation will be described in detail below. Note that the cell group aspirated from the grid 3G of the first well 3A1 may be discharged into a separately prepared container with a grid, rather than into the second well 3B1.

[0017] The destination plate 4 has a plurality of wells 41 into which the cells C picked from the sorting plate 2 are discharged. The wells 41 are holes with a bottom that open to the top surface of the destination plate 4. Each well 41 contains the required number of cells C (usually one) together with a liquid culture medium. The cells C contained in the well 41 are subjected to various tests such as the addition of reagents or reactants, as well as observation and culture. The destination plate 4 is also made of a member made of a translucent resin material or glass. For example, a commercially available 96-well plate (e.g., Corning Model No. 3595) can be used as the destination plate 4.

[0018] The camera unit 5 is a device that captures images of cells C held at the bottom of the sorting plate 2 or the destination plate 4 from the underside of these plates, and includes a lens unit 51 and a camera body 52. ​​The lens unit 51 is an objective lens used in optical microscopes and includes a group of lenses that form an optical image at a predetermined magnification and a lens barrel that houses this group of lenses. The camera body 52 includes an imaging element such as a CCD image sensor. The lens unit 51 forms an optical image of the object to be imaged on the light-receiving surface of the imaging element. The camera unit 5 is movable in the X and Y directions below the base 1 along guide rails 5G that extend left and right parallel to the base 1. The lens unit 51 is also movable in the Z direction for focusing. The camera unit 5 of this embodiment is capable of performing normal bright-field imaging and fluorescent imaging to capture fluorescent target cells C.

[0019] The head unit 6 is a device provided for picking cells C from the sorting plate 2 and moving them to the destination plate 4. The head unit 6 includes a plurality of heads 61 and a head body 62 to which these heads 61 are attached. A tip 10 having a tip portion 10T for aspirating and discharging cells C is attached to the tip of each head 61. The head body 62 holds the head 61 so that it can move up and down in the +Z and -Z directions (up and down directions), and can move in the +X and -X directions (horizontal directions) along guide rails 6G. The head body 62 can also move in the Y direction. In other words, the head 61 can move in three dimensions: X, Y, and Z.

[0020] The operation of the head 61 is roughly as follows: The head 61, to which the chip 10 is attached, is moved in the XY direction to a coordinate position corresponding to the position of the target cell C in the well 3 (first container) to be moved, which is identified based on the image captured by the camera unit 5. Next, the head 61 is lowered to approach the target cell C. Then, a cell group including the target cell C is aspirated into the chip 10. After aspirating, the head 61 is raised and moved in the XY direction to another well 3 (second container) in the sorting plate 2 or a separately prepared sorting container. After movement, the cell group held in the chip 10 is discharged into the other well 3 or sorting container. This dilutes cells other than the target cell C that are not to be moved. Next, only the target cell C is re-aspirated into the chip 10 from the well 3 or sorting container. Thereafter, the head 61 is moved in the XY direction to the destination plate 4 (third container), and the target cell C in the chip 10 is discharged into the specified well 41.

[0021] [Details of the Chip and Head] Figure 3(A) is a cross-sectional view of the chip 10 attached to the head 61, and a diagram showing the movement mechanism and suction mechanism of the chip 10. The chip 10 is a tool that sucks or discharges cells C in order to move the cells C, and is equipped with a tip portion 10T having a tip opening t through which the cells C can enter and exit. The chip 10 of this embodiment is composed of an assembly of a syringe 11 and a plunger 12. The syringe 11 has a tubular passage 11P therein, which serves as a suction path for the cells C. The plunger 12 moves back and forth within the tubular passage 11P while making sliding contact with the inner peripheral wall of the syringe 11 that defines the tubular passage 11P.

[0022] The syringe 11 includes a syringe base end 111 formed of a large-diameter cylindrical body and a syringe main body 112 formed of a long, thin-diameter cylindrical body. A tubular passage 11P is formed in the syringe main body 112. The tip opening t described above is provided in a syringe tip end 113 (tip end) at the lower end of the syringe main body 112. One end of the tubular passage 11P is connected to the tip opening t. The syringe base end 111 is connected to the other end of the syringe main body 112 via a tapered portion. The upper end portion of the syringe base end 111 is fitted into the lower end of the head 61.

[0023] The plunger 12 includes a cylindrical plunger base end 121, a needle-shaped plunger main body 122 continuing below the plunger base end 121, and a plunger tip end 123 which is the lower end of the plunger main body 122. The plunger 12 is assembled to the syringe 11 in such a manner that the plunger base end 121 is housed within the syringe base end 111 and the plunger main body 122 is inserted into the tubular passage 11P of the syringe main body 112. In the state shown in FIG. 3A where the plunger main body 122 is inserted to the deepest extent into the syringe main body 112, the plunger tip end 123 protrudes from the tip opening t. A rod 61R which is movable up and down within the internal space of the head 61 is attached to the upper end of the plunger base end 121.

[0024] A head driving unit 64 is provided inside the head main body 62. The head driving unit 64 functions as a mechanism for moving the tip 10 attached to the head 61 in the up and down directions, and as a mechanism for aspirating and discharging the cells C into the tip 10 through the tip opening t of the tip 10. The head driving unit 64 includes a head lifting motor 641 and a plunger lifting motor 642.

[0025] The head lift motor 641 is a motor that serves as a drive source for raising and lowering the head 61 relative to the head body 62. When the head 61 is raised and lowered by the driving of the head lift motor 641, the tip 10 attached to the lower end of the head 61 also rises and lowers. In other words, the height position of the tip opening t of the tip portion 10T can be set to a desired position by controlling the operation of the head lift motor 641.

[0026] The plunger lift motor 642 is a motor that serves as a drive source for raising and lowering the rod 61R within the internal space of the head 61. When the rod 61R is raised and lowered, the plunger 12 attached to this rod 61R also rises and lowers. When the plunger 12 rises relative to the syringe 11, a suction force is generated at the tip opening t. On the other hand, when the plunger 12 descends relative to the syringe 11, a discharge force is generated at the tip opening t. In other words, by controlling the operation of the plunger lift motor 642, the suction operation and discharge operation of the cell C by the tip 10 can be controlled.

[0027] Fig. 3(A) shows the state in which the plunger 12 is lowered to the lowest position. This state is the state before cells C are aspirated, or the state in which cells C aspirated into the tip 10 have been discharged. The plunger tip 123 protrudes slightly downward from the syringe tip 113. Fig. 3(B) shows the state in which the plunger 12 is raised by a predetermined height. This state is the state of the tip 10 during the aspirating operation to aspirate cells C. Fig. 3(C) shows an enlarged view of the main parts of Fig. 3(B).

[0028] During the suction operation, the plunger tip 123 sinks into the tubular passage 11P. At this time, a suction force is generated at the tip opening t, and the fluid around the tip opening t is sucked into the suction space H formed in the tubular passage 11P by the plunger tip 123 sinking inward. In other words, the culture medium LCM containing the cells C is held in the suction space H. After this suction operation, when the plunger 12 is moved downward, the fluid held in the suction space H is discharged from the tip opening t. The amount of fluid sucked can be adjusted by the height to which the plunger 12 rises, and the speed at which the fluid is sucked can be adjusted by the rate at which the plunger 12 rises.

[0029] [Electrical Configuration of Cell Migration Device] Figure 4 is a block diagram showing the electrical configuration of the cell migration device S. The cell migration device S is equipped with a control unit 7 that controls the movement of the head unit 6 (see Figure 1) and the elevation of the head 61 (chip 10), i.e., the three-dimensional movement of the head 61. In addition, the control unit 7 controls the suction and discharge of cells C into the chip 10, as well as the movement and image capture of the camera unit 5.

[0030] The cell movement device S also includes a camera axis drive unit 53, a servo motor 54, a head unit axis drive unit 63, and a head drive unit 64. The camera axis drive unit 53 includes a drive motor that moves the camera unit 5 horizontally along the guide rail 5G (FIG. 1). The servo motor 54 rotates forward or backward to move the lens unit 51 vertically at a predetermined resolution via a power transmission mechanism (not shown). This allows the focal position of the lens unit 51 to be adjusted to the cell C contained in the well 3. As shown by the dotted line in FIG. 4, the base 1 may be moved vertically instead of the lens unit 51. The head unit axis drive unit 63 includes a drive motor that moves the head unit 6 (head main body 62) horizontally in the X or Y direction along the guide rail 6G. The head drive unit 64 is as described above with reference to FIG. 3.

[0031] The control unit 7 is made up of a processor and the like, and by executing a predetermined program, functions to include an axis control unit 71, a head control unit 72, an imaging control unit 73, an image processing unit 74, a memory unit 75, and a main control unit 78. Furthermore, the control unit 7 is provided with an input unit 76 for inputting various information to the control unit 7, and a display unit 77 for displaying various information. The input unit 76 accepts input of various operational information from the operator. The display unit 77 functions as a monitor for displaying images captured by the camera unit 5, etc.

[0032] The axis control unit 71 controls the operation of the head unit axis drive unit 63. By controlling the head unit axis drive unit 63, the axis control unit 71 moves the head unit 6 to a predetermined target position in the horizontal direction. Movement of the head 61 (chip 10) between the sorting plate 2 and the destination plate 4 and between the wells 3, positioning vertically above the cells C contained in the wells 3, and positioning vertically above the wells 41 to be discharged are achieved by the axis control unit 71 controlling the head unit axis drive unit 63. The axis control unit 71 also controls the camera axis drive unit 53 to control the operation of moving the camera unit 5 along the guide rails 5G.

[0033] The head control unit 72 raises and lowers the head 61 to be controlled toward a predetermined target position by controlling the head lift motor 641 of the head drive unit 64. The head control unit 72 also controls the plunger lift motor 642 to generate a suction force or a discharge force at the tip opening t of the tip 10 at a predetermined timing.

[0034] The imaging control unit 73 controls the imaging operation of the camera unit 5 of the selection plate 2 or the destination plate 4, such as the exposure amount and shutter timing. For focusing, the imaging control unit 73 also provides control pulses to the servo motor 54 to move the lens unit 51 vertically at a predetermined pitch (for example, a pitch of several tens of μm). The imaging control unit 73 can cause the camera unit 5 to perform normal bright-field imaging and fluorescent imaging in which the target cells C fluoresce.

[0035] The image processing unit 74 performs image processing such as edge detection and pattern recognition processing involving feature extraction on the image data acquired by the camera body 52. ​​Based on an image of the sorting plate 2 containing the cells C, the image processing unit 74 performs processing such as recognizing the presence of the cells C on the bottom surface 31 of the well 3 and the state of the cells C contained in the grid 3G (plurality of recesses) on the image. Similarly, based on an image of the well 41 to which the cells C have been transferred, the image processing unit 74 performs processing such as recognizing the number, amount, fluorescence intensity, etc. of the cells C contained in the well 41.

[0036] The storage unit 75 stores various setting values, data, programs, etc. for the cell migration device S. In addition, the storage unit 75 stores information about the sorting plate 2 to be used, such as the plate size, the size of the wells 3, and the size of the grid 3G. Information about the tip shape of the tip 10, such as the outer diameter, thickness, and opening diameter of the tip opening t of the syringe tip 113, is also stored in the storage unit 75. Furthermore, setting information such as the suction amount and suction speed in the suction operation of the cells C is also stored in the storage unit 75.

[0037] The main control unit 78 comprehensively controls the operations of the camera unit 5 and the head unit 6. The main control unit 78 captures images of the sorting plate 2 and controls a picking operation in which target cells C in wells 3 selected as the movement target are aspirated into the tip 10 attached to the head 61. The main control unit 78 also controls a moving operation in which the aspirated target cells C are moved to other wells 3 in the sorting plate 2 or to the destination plate 4, as well as a discharging operation in which the aspirated target cells C are discharged. The main control unit 78 controls the camera unit 5 and the head unit 6 via the axis control unit 71, the head control unit 72, and the imaging control unit 73 to perform the above-mentioned picking operation, moving operation, and discharging operation.

[0038] For the above control, the main control unit 78 functionally includes a movement and discharge control unit 781, a discharge pressure control unit 782, and a picking control unit 783. The movement and discharge control unit 781 controls the cell discharge operation from the chip 10 and the movement operation of the head 61 during discharge. The discharge pressure control unit 782 controls the discharge pressure during cell discharge from the chip 10, i.e., the positive pressure for discharge applied to the tip opening t. The picking control unit 783 controls the picking operation of the cell C by the chip 10 through the axis control unit 71 and the head control unit 72.

[0039] [Cell Discharge Operation According to the Present Embodiment] When discharging a large number of cells C into a well 3 having a grid 3G on the bottom surface 31 as illustrated in FIG. 2(B), it is desirable to discharge the cells C so that they are dispersed and accommodated on as many grids 3G as possible. If the cells C are unevenly arranged on the bottom surface 31, it becomes difficult to selectively pick the target cells C to be moved. Ideally, one cell C is accommodated on one grid 3G, that is, the cell C is isolated. In this case, only the target cells C can be easily picked by the chip 10. In this embodiment, the cells C are discharged from the chip 10 so that the target cells C can be picked efficiently after the cells are discharged.

[0040] 5 is a cross-sectional view showing the state of cell dispensing from the chip 10 to the well 3. The movement and dispensing control unit 781 controls the dispensing of cells C from the chip 10 while moving the head 61. When dispensing cells, liquid culture medium LCM is stored in advance in the well 3. In other words, the grid 3G and the space above it are filled with liquid culture medium LCM. Meanwhile, a plurality of cells C aspirated from a separate container are held in the suction space H of the chip 10. The movement and dispensing control unit 781 controls the axis control unit 71 and head control unit 72 to move the head 61 in the XYZ directions, and immerses the tip 10T of the chip 10 in the liquid culture medium LCM in the well 3.

[0041] After the above state is achieved, the movement and discharge control unit 781 discharges cells C from the tip opening t of the tip 10 while moving the head 61. That is, while the tip 10 is moved in the XY direction as shown by arrow a1 or arrow a2 in conjunction with the XY movement of the head 61, the plunger main body 122 is lowered to discharge the cells C held in the suction space H from the tip opening t. The discharged cells C settle under their own weight while spreading as the head 61 moves, and are eventually accommodated in the grid 3G on the bottom surface 31. In this way, the cells C are discharged from the tip 10 while moving the tip 10. This allows the cells C to be distributed in a scattered manner without being concentrated on a specific part of the bottom surface 31 of the well 3. This makes it easier to isolate the target cells C to be moved.

[0042] The manner of movement of the head 61 during discharging of cells C may be set as appropriate as long as it promotes cell dispersion. Figures 6A to 6E are diagrams showing various examples of movement of the chip 10 during cell discharging. Figure 6A shows a manner in which the head 61 moves linearly in the X or Y direction. The head 61 may be reciprocated in the X or Y direction, or may be moved diagonally by simultaneously moving in the X and Y directions. Furthermore, the head 61 may be moved linearly in the X or Y direction while slightly reciprocating in the Z direction, i.e., raised and lowered. Figure 6B shows an example of moving the head 61 in a zigzag pattern. For example, the head 61 may be moved linearly in the X direction while reciprocating (oscillating) in the Y direction, or moved linearly in the X or Y direction while moving in the Z direction.

[0043] 6(C) shows an example in which the head 61 moves in a circular spiral in a plan view, and FIG. 6(D) shows an example in which the head 61 moves in a rectangular spiral. The starting point of the spiral movement may be either the center position or the outermost position of the spiral. However, since using the outermost periphery as the starting point is likely to cause interference between the chip 10 and the sidewall of the well 3, it is preferable to use the center position as the starting point. Note that instead of a spiral movement, the head 61 may simply move along a circular or elliptical orbit, or a square or rectangular orbit.

[0044] 6(E) shows an example in which the head 61 is vibrated. The vibration can be achieved by reciprocating the head 61 over a small distance in the X or Y direction. The head 61 may be vibrated in the Y direction while moving straight in the X direction, or may be vibrated in the X direction while moving straight in the Y direction. Alternatively, the head 61 may be positioned at a fixed point in the well 3 and vibrated for a predetermined time, and then the fixed point may be moved to another position in the well 3 and vibrated for a predetermined time. In addition to vibration in the X and Y directions, the head 61 may be vibrated in the Z direction.

[0045] [Example of Picking Target Cells from a Large Number of Cells] When the number of target cells is very small relative to the total number of cells contained in a single container, it is difficult to selectively pick the target cells. Conventionally, limiting dilution is applied to dilute a cell suspension containing target cells before picking. However, limiting dilution requires the dispensing of cell-containing culture medium into wells. Therefore, when the total number of cells is large, multiple dispensing operations are required, which is extremely time-consuming and requires a large number of well plates. In view of this, this embodiment shows an example in which target cells can be efficiently isolated from a large number of cells and easily picked.

[0046] 7A to 7D are cross-sectional views illustrating the process from seeding cells C to picking target cells Ct using the first well 3A1 (first container) and second well 3B1 (second container) of the well 3 illustrated in FIG. 2. The first well 3A1 includes a first bottom surface 31A having a plurality of first grids 3G1 (first recesses). In FIG. 7A, five grids 3G11, 3G12, 3G13, 3G14, and 3G15 are shown as the first grids 3G1. The second well 3B1 includes a second bottom surface 31B having a plurality of second grids 3G2 (second recesses). In FIG. 7C, five grids 3G21, 3G22, 3G23, 3G24, and 3G25 are shown as the second grids 3G2. A predetermined amount of liquid medium LCM is stored in the first well 3A1 and the second well 3B1.

[0047] 7(A) to 7(D) roughly consist of the following steps (1) to (5): (1) seeding a large amount of cells into the first well 3A1, (2) aspirating a plurality of cells C from the first well 3A1 into the chip 10, (3) moving the chip 10 holding the cells C to the second well 3B1, (4) discharging the cells C while moving the chip 10 with the tip 10T of the chip 10 immersed in the liquid medium LCM in the second well 3B1, and (5) picking the target cells Ct from the second well 3B1.

[0048] FIG. 7(A) shows the implementation of step (1) above. The chip 10 faces the first well 3A1 in order to seed a large number of cells C. The chip 10 holds a large number of cells C previously aspirated from a dispensing tube. When seeding the cells C in the first well 3A1, the chip 10 is moved while discharging the cells to improve the dispersibility of the cells C. The movement and discharge control unit 781 (FIG. 4) discharges the cells C from the chip 10 while moving the head 61 with the tip 10T of the chip 10 immersed in the liquid culture medium LCM in the first well 3A1. Here, an example in which the chip 10 is moved horizontally is shown.

[0049] The seeding of cells C is performed so that a predetermined number of cells C is accommodated in each of the first grids 3G1 provided in the first well 3A1. As an example, assume that the first well 3A1 has 16,000 square first grids 3G1 with sides of 200 μm. In this case, cells C are seeded in an amount such that 150 cells C are accommodated per first grid 3G1. That is, 150 × 16,000 = 2.4 million cells C are seeded in the first well 3A1. If the entire amount cannot be seeded in a single discharge operation from the chip 10, the seeding operation shown in FIG. 7A is repeated. If the target cell Ct to be picked is present at a rate of only one per 10,000 cells, the above example would contain 240 target cells Ct.

[0050] A target cell Ct is identified from among the cells C seeded in the first well 3A1. Examples of the means for identifying the target cell Ct include, for example, exciting the target cell Ct with fluorescence or binding a fluorescent protein to the target cell Ct, and then capturing an image of the fluorescence with the camera unit 5. Figure 7(A) shows an example in which a cell C is housed in each of the grids 3G11 to 3G15, and only the target cell Ct is contained in grid 3G13. The picking control unit 783 identifies grid 3G13, which houses the target cell Ct, as the target grid (target recess) to be aspirated in step (2) above.

[0051] 7(B) shows the implementation status of step (2). The picking control unit 783 aligns the head 61 with a new chip 10 attached to the target grid 3G13 and causes the chip 10 to aspirate all of the cells C present on the target grid 3G13. This suction causes the chip 10 to retain the target cells Ct and cells C that are not to be moved. In the above example, statistical calculations indicate that 150 cells C are aspirated into the chip 10, one of which is the target cell Ct. After the suction is complete, the head 61 is moved so that the chip 10 moves toward the second well 3B1, as in step (3) above.

[0052] 7(C) shows the implementation of step (4). The movement and discharge control unit 781 discharges cells C while moving the tip 10 with the tip 10's tip end 10T immersed in the liquid culture medium LCM in the second well 3B1. The cells C are seeded into the second well 3B1 so that a desired number of cells C are dispersedly accommodated in each of the second grids 3G2 provided in the second well 3B1. The ideal dispersion state is one cell C accommodated in each second grid 3G2. Step (4) may be repeated as long as such a dispersion state can be ensured.

[0053] Here is a specific example. Similar to the first well 3A1 exemplified above, the second well 3B1 also has 16,000 square first grids 3G1 with sides of 200 μm. In this case, even if approximately 4,000 to 8,000 cells C are seeded, the probability of occurrence of a second grid 3G2 containing only one cell C can be maintained high. In the above example, 150 cells are picked by suction from one target grid 3G13, so steps (2) to (4) can be repeated approximately 25 to 55 times.

[0054] The state of the cell C contained in the second well 3B1 is confirmed based on the image acquired by the camera unit 5. Figure 7(C) shows an example in which one target cell Ct is contained in grid 3G23 of grids 3G21 to 3G25 of the second well 3B1. The containment position of the target cell Ct is identified based on the results of fluorescent imaging by the camera unit 5. The picking control unit 783 identifies grid 3G23 containing the target cell Ct as the target grid.

[0055] 7(D) shows the implementation status of step (5). The picking control unit 783 aligns the head 61 equipped with a new chip 10 with the target grid 3G23 and causes the chip 10 to aspirate the isolated target cells Ct. After aspirating, the head 61 is moved so that the chip 10 moves toward the destination plate 4. After movement, the target cells Ct are ejected from the chip 10 into one well 41 of the destination plate 4. Step (5) is repeated the number of times equal to the number of target cells Ct isolated in the second well 3B1.

[0056] According to this embodiment, a target grid 3G13 containing target cells Ct is identified from among the first grids 3G1 in the first well 3A1, and all cells C present on that target grid 3G13 are aspirated. The cell group containing the aspirated target cells Ct is then ejected into the second well 3B1. In both cases, cells are ejected while the chip 10 is moving, dispersing the cells C and allowing the cells C to be evenly distributed across multiple grids 3G1 and 3G2. Furthermore, the above two-step ejection naturally dilutes cells not targeted for movement. In the specific example described above, while one target cell Ct was found among 10,000 cells C in the first well 3A1, one target cell Ct was found among 150 cells C in the second well 3B1. This facilitates isolation of target cells Ct in the second well 3B1, facilitating subsequent picking.

[0057] [Various ideas for cell ejection] The above is an example of the process from seeding cell C to picking target cell Ct, but the preferred ejection mode for cell ejection in the above step (1) (FIG. 7(A)) and step (4) (FIG. 7(C)) is shown below.

[0058] <Setting the Discharge Pressure> For example, when discharging target cells Ct from the chip 10 into the wells 41 of the destination plate 4, the discharge pressure controller 782 controls the application of a predetermined reference positive pressure for discharge to the tip opening t of the chip 10. Specifically, the discharge pressure controller 782 controls the descending speed of the plunger 12 ( FIG. 3 ). The reference positive pressure is set to a pressure at which the cells C held in the suction space H of the chip 10 are quickly discharged from the tip opening t.

[0059] On the other hand, in the above steps (1) and (4), rapid discharge can impair the dispersion of cells C. This is because applying a standard positive pressure for discharge to the tip opening t of the chip 10 causes cells C held within the chip 10 to suddenly and forcefully eject. Therefore, when discharging cells while moving the chip 10, positive pressure may not be applied to the tip opening t. That is, the head 61 may be moved without applying positive pressure for discharge to the tip opening t while the tip 10T of the chip 10 is immersed in the liquid medium LCM in the first well 3A1 or the second well 3B1. According to this embodiment, cells C in the chip 10 fall freely from the tip opening t toward the bottom surfaces 31A and 31B by gravity. Furthermore, the chip 10 is moving. This prevents cells C from expelling, making it easier to disperse the cells C and distribute them evenly across the grids 3G1 and 3G2 on the bottom surfaces 31A and 31B.

[0060] If the process relies solely on the free fall, it may take time to eject the cells C held in the chip 10. Therefore, a positive pressure smaller than the reference positive pressure may be applied to the tip opening t during steps (1) and (4). For example, a positive pressure of about 1 / 2 to 1 / 10 of the reference positive pressure may be applied to the tip opening t. This embodiment can reduce the time required to eject the cells while preventing the cells C from flying out of the chip 10 with force.

[0061] <Dispersion of cells within the chip> Cells C settle within the liquid medium LCM. For this reason, cells C aspirated into the chip 10 together with the liquid medium LCM will settle near the tip 10T if time is required between aspiration and discharge. If discharge pressure is applied to the tip opening t in this state, cells C unevenly distributed near the tip 10T may be released all at once, impairing dispersibility. In anticipation of this case, it is desirable to intervene a step of dispersing cells C within the chip 10.

[0062] 8A to 8D are diagrams showing an example of seeding cells C into wells 3, including a step of dispersing cells within the chip 10. Here, the first seeding of step (1) above is assumed. As shown in FIG. 8A, the tip 10T of the chip 10 is inserted into a dispensing tube 21 that stores a cell suspension containing a large number of cells C in a liquid medium LCM. Then, a large number of cells C are aspirated into the chip 10 together with the liquid medium LCM.

[0063] Next, as shown in FIG. 8(B), the head 61 is raised to lift the chip 10, and the head 61 is moved toward the well 3 where seeding will be performed. At this time, the cells C aspirated into the chip 10 settle near the tip 10T. If the tip 10T is then placed in the liquid medium LCM in the well 3 in this state, the cells C may unintentionally leak out all at once from the tip opening t. Therefore, as shown in FIG. 8(C), a slight suction force is generated in the chip 10, and the cells C that have formed clumps near the tip 10T are lifted upward and dispersed. In other words, the chip 10 is caused to perform a suction operation so that the cells C aspirated into the chip 10 move toward the depth of the chip 10.

[0064] 8(D), the cells C are ejected while the tip 10 is moved with the tip 10T immersed in the liquid medium LCM in the well 3. Before the horizontal movement and ejection of the chip 10 are actually performed, as shown in step (D1), a slight suction force is generated in the chip 10, causing the chip 10 to suck in a small amount of the liquid medium LCM, thereby causing the cells C in the chip 10 to soar. In other words, this operation of causing the chip 10 to perform a suction operation so that the cells C sucked into the chip 10 move toward the depth of the chip 10 further disperses the cells C in the chip 10.

[0065] In the subsequent steps (D2) and (D3), cells C are discharged while the chip 10 is being moved. Because the cells C are dispersed within the chip 10, a large amount of cells C is not discharged all at once. The discharged cells C settle and are accommodated in the grid 3G on the bottom surface 31. Between steps (D2) to (D3), a step of re-aspirating a small amount of liquid culture medium LCM into the chip 10, similar to step (D1), may be inserted. This allows the cells C that are settling again within the chip 10 to be lifted up and dispersed again.

[0066] <Detection of Cell-Undispersed Region> When cells C are seeded into well 3, it is desirable that the cells C are dispersed evenly across the entire area of ​​bottom surface 31. If there is an undispersed region on bottom surface 31 where cells C are not contained, it is desirable that cells C be ejected from chip 10 toward that undispersed region. As shown as an example in FIG. 7(C) and in step (4) above, when cells C are seeded into second well 3B1, cells are ejected from chip 10 multiple times. It is desirable to identify an undispersed region during multiple cell ejections, and then eject cells targeting that undispersed region.

[0067] 9A to 9C are diagrams showing an example of cell discharge that ultimately prevents the formation of cell-undispersed regions on the bottom surface 31 of the well 3. Fig. 9A shows a state in which cell seeding has progressed to a certain extent and cells C are dispersed on the bottom surface 31. The dispersion state of cells C on the bottom surface 31 is detected by the image processing unit 74 performing image processing such as object detection processing and filtering processing on the image captured by the camera unit 5.

[0068] FIG. 9(B) is a diagram showing an example of image processing. Here, an image is shown in which cells C that are contained in only one grid 3G and cells C that are contained in two or more grids 3G are color-coded. White cells C are isolated cells C. If a target cell Ct is present among the white cells C, it will be picked. The image in FIG. 9(B) also clearly shows undispersed regions MA where cells C have not yet been dispersed.

[0069] 9(C) shows an example of setting a movement line 10L of the chip 10 when subsequent cell ejection is performed on the detected undispersed region MA. The movement line 10L is set so that the chip 10 passes through the undispersed region MA. The movement and ejection control unit 781 ejects cells C from the chip 10 while moving the head 61 along the movement line 10L. This causes the cells to be ejected toward the undispersed region MA of the cells C on the bottom surface 31, allowing the cells C to be evenly distributed across the entire area of ​​the bottom surface 31.

[0070] [Cell Picking Control Flow] Figure 10 is a flowchart showing the cell picking process executed by the main controller 78. The "first container" and "second container" shown in Figure 10 are independent containers or two wells on a single plate. In the following, following the example described above, the "first well 3A1" of the sorting plate 2 will be referred to as the "first container" and the "second well 3B1" as the "second container."

[0071] The movement / discharge control unit 781 of the main control unit 78 controls the axis control unit 71 and the head control unit 72 to operate the head 61, and seed sample cells to be picked from the chip 10 attached to the head 61 into the first well 3A1 of the sorting plate 2 (step S1). The seeding operation is an operation in which the head 61 is moved horizontally and cells C are discharged from the chip 10, as described in Fig. 7(A).

[0072] Next, the imaging control unit 73 controls the camera unit 5 to capture an image of the first bottom surface 31A of the first well 3A1, and the placement state of the seeded cells C on the grid 3G is confirmed on the image (step S2). Bright-field imaging and fluorescent imaging, which causes the target cells Ct to fluoresce, are performed as the imaging. Phase-contrast imaging may also be performed as the imaging. The placement state of all cells on the grid 3G is confirmed based on the image acquired by bright-field imaging. The presence of the target cells Ct is confirmed based on the image acquired by fluorescent imaging. This confirmation operation can be performed by image processing by the image processing unit 74, which recognizes points of high brightness in the image. Of course, the image may also be displayed on the display unit 77 for visual confirmation by the operator.

[0073] Next, it is determined whether or not fluorescent cells, i.e., target cells Ct, have been recognized in the image captured in step S2 (step S3). If target cells Ct have not been recognized (NO in step S3), it is determined that target cells Ct do not exist (step S4), and a message to that effect is displayed on the display unit 77. In this case, the operator must either reseede the first well 3A1 or seed another well of the selection plate 2.

[0074] If the target cell Ct is recognized (YES in step S3), the selection of the first grid 3G1 containing the target cell Ct is accepted (step S5). That is, the selection of the target cell Ct to be moved to the second well 3B1 is accepted as the selection of the first grid 3G1. For example, the fluorescent image is displayed on the display unit 77, and the operator's grid selection operation is accepted via the input unit 76. In the example of FIG. 7(A), grid 3G13 is selected. Note that the selection of the grid containing the target cell Ct may be performed automatically based on the image recognition results.

[0075] Next, the picking control unit 783 controls the axis control unit 71 and the head control unit 72 to operate the head 61, causing all of the cells C contained in the first grid 3G1 selected in step S5 to be aspirated into the chip 10 (step S6). This operation is as shown in FIG. 7(B). Thereafter, the axis control unit 71 moves the head 61 to above the second well 3B1 (step S7). In other words, the chip 10 holding the cell group including the target cells Ct is moved to the second well 3B1.

[0076] Next, the movement and discharge control unit 781 operates the head 61 to immerse the tip 10T of the chip 10 into the liquid culture medium LCM stored in the second well 3B1 (step S8). Then, as illustrated in Fig. 7(C), the movement and discharge control unit 781 operates the head 61 to horizontally move the chip 10 and discharge a cell group including the target cells Ct from the chip 10 (step S9).

[0077] Thereafter, it is confirmed whether a predetermined amount of cells C has been dispensed into the second well 3B1 (step S10). In the above example, approximately 4,000 to 8,000 cells C can be seeded into the second well 3B1. In step S10, it is confirmed whether seeding of this number of cells has been completed. If the predetermined amount of cells C has not been dispensed (NO in step S10), it is confirmed whether the grid 3G1 selected in step S5, i.e., the grid containing the target cells Ct, remains (step S11).

[0078] If any selection grids containing target cells Ct remain (YES in step S11), the process returns to step S6 and is repeated. On the other hand, if no selection grids remain (NO in step S11), the seeding is terminated even if there is room for additional seeding in the second well 3B1. Also, if the predetermined amount of cells C has been dispensed into the second well 3B1 (YES in step S10), the seeding is terminated.

[0079] Thereafter, the second bottom surface 31B of the second well 3B1 is imaged by the camera unit 5, and the state of the seeded cells C contained in the second grid 3G2 is confirmed on the image (step S12). Here, as in step S2, bright-field imaging and fluorescent imaging, which causes the target cells Ct to fluoresce, are performed. Once the target cells Ct isolated on the second grid 3G2 are confirmed, the picking control unit 783 performs an operation of aspirating the target cells Ct into the chip 10 (step S13).

[0080] [Inventions Included in the Above-Described Embodiments] The above-described embodiments include the following inventions.

[0081] A cell movement device according to one aspect of the present invention is a cell movement device that moves a plurality of cells contained in a first container to a second container that stores a liquid, and is equipped with a head that is equipped with a tip having a tip that aspirates and ejects cells, and that aspirates a plurality of cells in the first container into the tip, moves them to the second container, and ejects the plurality of cells in the tip into the second container, and a control unit that controls the operation of the head, and the control unit controls the head to eject cells from the tip while moving the head with the tip of the tip immersed in the liquid in the second container.

[0082] According to this cell transfer device, when discharging a plurality of cells sucked into the tip into the second container, the head is moved. In other words, the cells are discharged from the tip while the tip is being moved. This allows the cells to be distributed in a scattered manner rather than being concentrated in one part of the second container. This makes it easier to isolate the target cells to be transferred, and facilitates the picking of the target cells.

[0083] In the above-mentioned cell movement device, the first container may be a container for storing liquid, and the control unit may eject cells from the tip while moving the head, with the tip of the tip, which has previously aspirated a large number of cells, immersed in the liquid in the first container.

[0084] According to this aspect, when the cells are seeded into the first container, the cells are discharged from the tip while the tip is being moved, so that the cells can be dispersed evenly into the first container.

[0085] In the above-mentioned cell movement device, the second container includes a bottom having a plurality of recesses, and the control unit controls the movement of the head and the ejection so that the desired number of cells are dispersedly contained in the plurality of recesses.

[0086] According to this embodiment, it is possible to prevent the cells from being unevenly accommodated in a particular well. For example, by applying a statistical method to adjust the number of cells to be seeded, taking into account the bottom area and the number of wells, it is possible to roughly adjust the number of cells accommodated in each well.

[0087] In the above-mentioned cell movement device, it is desirable that the first container includes a first bottom having a plurality of first recesses, and the second container includes a second bottom having a plurality of second recesses, and after discharging cells into the first container, the control unit identifies a target recess among the plurality of first recesses that contains target cells to be moved, causes all of the cells present in the target recess to be sucked into the tip, and moves the head to the second container, and performs the movement of the head and the discharge so that the desired number of cells are dispersedly contained in the plurality of second recesses.

[0088] According to this aspect, a target well is identified from among a plurality of first wells, and all cells present in that target well are aspirated. Next, the cell population containing the aspirated target cells is discharged into a second container. These two steps naturally dilute the concentration of non-target cells relative to the target cells. For example, if one target cell is contained in 10,000 cells in the first container, it is possible to create a state in which one target cell is contained in 100 cells in the second container. This facilitates picking of target cells from the second container, thereby improving the efficiency of the picking operation.

[0089] In the cell migration device, the control unit can move the head so as to trace a spiral trajectory in a plan view.

[0090] According to this embodiment, the tip that discharges the cells can be easily passed through the entire cavity of the first container or the second container evenly, and therefore the cells can be easily dispersed over the entire bottom surface of the container.

[0091] In the cell migration device, the control unit may further vibrate the head while moving the head, thereby making it easier to disperse the cells.

[0092] In the above-mentioned cell movement device, the control unit may move the head without applying positive pressure for ejection to the tip of the tip while the tip of the tip is immersed in the liquid in the second container.

[0093] When positive pressure for discharging is applied to the tip of the tip, cells sucked into the tip may be ejected with force. According to the above embodiment, cells can be placed on the bottom of the container by free fall using gravity. Therefore, the cells can be easily dispersed and placed on the bottom of the container. Here, "placement" includes adhering the cells to the bottom of the container and floating them on the bottom of the container.

[0094] In the above-mentioned cell movement device, the positive pressure applied to the chip holding a single cell when ejecting the cell from the chip is predetermined as a reference positive pressure, and the control unit moves the head by applying a positive pressure smaller than the reference positive pressure to the tip of the chip while the tip of the chip is immersed in the liquid in the second container.

[0095] According to this embodiment, cells can be ejected more quickly than when cells are ejected from the tip by free fall. Furthermore, since the ejection pressure is a positive pressure that is smaller than the reference positive pressure, cells can be prevented from flying out of the tip with force.

[0096] In the above-mentioned cell movement device, the control unit may immerse the tip of the chip in the liquid of the second container, cause the chip to perform a suction operation so that the multiple cells sucked into the chip move toward the depth of the chip, and then eject the cells from the chip while moving the head.

[0097] According to this aspect, by performing the suction operation before discharging the cells, the cells sucked into the tip are dispersed, thereby preventing a large number of cells from being released from the tip all at once when the cells are discharged.

[0098] In the above-mentioned cell movement device, the control unit may cause the tip to perform a suction operation so that the multiple cells sucked into the tip move toward the depth of the tip before immersing the tip in the liquid in the second container.

[0099] According to this aspect, the suction operation is performed before the tip of the tip is immersed in the liquid in the second container. Therefore, even if cells aspirated to the tip of the tip settle, these cells can be lifted upward. Therefore, cells can be prevented from falling from the tip of the tip into the liquid in the second container at the beginning of the tip immersion. In addition, since the cells are dispersed within the tip before discharge, there is also the advantage that the amount of cells discharged can be easily adjusted.

[0100] In the above-mentioned cell movement device, it is desirable that the device further includes a camera capable of capturing an image of the bottom of the first container or the second container, and that the control unit has an image processing unit that identifies the state of cell storage in the multiple recesses based on the image captured by the camera.

[0101] According to this aspect, the state of cell dispersion at the bottom of the first container or the second container and the position of the target cell can be easily identified based on the camera image.

[0102] In the above-mentioned cell movement device, the image processing unit may identify an undispersed region at the bottom of the second container that does not contain cells based on the image, and the control unit may eject cells from the tip while moving the head so that the tip passes through the undispersed region.

[0103] According to this aspect, the cells are discharged toward the undispersed cell region at the bottom of the second container, thereby allowing the cells to be distributed evenly over the entire area of ​​the bottom.

[0104] The above-mentioned cell movement device may further include a third container into which target cells are moved from the second container, and the control unit may identify a second container containing target cells from among the plurality of second containers, suck the target cells from the second container into the tip, and move the head to the third container and eject the target cells from the tip into the third container.

[0105] According to this embodiment, only the target cells can be picked and transferred to the third container. By transferring the cells from the first container to the second container, the ratio of target cells to all cells is increased. Therefore, the probability of a second well containing a single target cell also increases. Therefore, it is easy to identify a second well containing a target cell from among multiple second wells and pick it.

[0106] In the cell migration device, it is desirable that a plurality of heads are provided, and that the control unit be capable of simultaneously or individually controlling the operation of the plurality of heads.

[0107] According to this embodiment, it is possible to use a plurality of heads and the chips attached to each of these, which means that the plurality of chips can be used simultaneously or individually, thereby improving the efficiency of the cell picking and transfer operations.

[0108] Another aspect of the present invention is a cell transfer method for transferring a plurality of cells contained in a first container to a second container that stores a liquid, in which a tip having a tip for aspirating and discharging cells is used to aspirate a plurality of cells into the tip in the first container, the tip is moved to the second container, and the cells are discharged from the tip while the tip is moved with the tip immersed in the liquid in the second container.

[0109] According to this cell transfer method, when a plurality of cells aspirated into a tip are discharged into a second container, the cells are discharged from the tip while the tip is being moved. This allows the cells to be dispersed in the second container without being concentrated in a certain area. This makes it easier to isolate the target cells to be transferred, and facilitates the picking of the target cells.

[0110] In the above-described cell transfer method, it is desirable that the first container includes a first bottom portion that stores a liquid and has a plurality of first recesses, and the second container includes a second bottom portion that has a plurality of second recesses, and while the tip of the tip that has previously aspirated a large number of cells is immersed in the liquid in the first container, the tip is moved while discharging the cells from the tip, a target recess among the plurality of first recesses that contains target cells to be transferred is identified, all of the cells present in the target recess are aspirated into the tip, and the tip is moved to the second container, and after the discharging into the second container, the state of cells contained in the plurality of second recesses is confirmed, the second recess in which the target cells are contained is identified, and the target cells are aspirated from the identified second recess with the tip.

[0111] According to this aspect, a target well is identified from among the plurality of first wells, and all cells present in that target well are aspirated. Next, a cell population containing the aspirated target cells is discharged into a second container. These two steps naturally dilute the concentration of non-target cells relative to the target cells. Therefore, picking of target cells from the second container can be easily performed, improving the efficiency of the picking operation.

[0112] According to the present invention, it is possible to provide a cell migration device and method that can efficiently pick a target cell from among a large number of cells.

Claims

1. A cell movement device that moves a plurality of cells contained in a first container to a second container that stores liquid, comprising: a head equipped with a tip having a tip for aspirating and discharging cells, and performing operations of aspirating a plurality of cells in the first container into the tip, moving the cells to the second container, and discharging the plurality of cells in the tip into the second container; and a control unit that controls the operation of the head, wherein the control unit controls the head to discharge cells from the tip while moving the head with the tip of the tip immersed in the liquid in the second container.

2. A cell movement device as described in claim 1, wherein the first container is a container for storing liquid, and the control unit ejects cells from the tip while moving the head, with the tip of the tip, which has previously aspirated a large number of cells, immersed in the liquid in the first container.

3. A cell movement device as described in claim 1, wherein the second container includes a bottom having a plurality of recesses, and the control unit controls the movement of the head and the ejection so that a desired number of cells are dispersed and contained in the plurality of recesses.

4. A cell movement device as described in claim 2, wherein the first container includes a first bottom having a plurality of first recesses, and the second container includes a second bottom having a plurality of second recesses, and the control unit, after discharging cells into the first container, identifies a target recess among the plurality of first recesses that contains a target cell to be moved, causes all of the cells present in the target recess to be sucked into the tip and moves the head to the second container, and causes the head to move and the discharge to be performed so that a desired number of cells are dispersed and contained in the plurality of second recesses.

5. A cell migration device according to any one of claims 1 to 4, wherein the control unit moves the head so as to trace a spiral trajectory in a planar view.

6. A cell migration device according to any one of claims 1 to 4, wherein the control unit further applies vibration to the head while causing the head to move.

7. A cell migration device as described in any one of claims 1 to 4, wherein the control unit moves the head while the tip of the tip is immersed in the liquid in the second container, without applying positive pressure for ejection to the tip of the tip.

8. A cell movement device according to any one of claims 1 to 4, wherein a positive pressure applied to the tip holding a single cell when ejecting the cell from the tip is determined in advance as a reference positive pressure, and the control unit applies a positive pressure smaller than the reference positive pressure to the tip of the tip while the tip is immersed in the liquid in the second container, thereby moving the head.

9. A cell movement device as described in any one of claims 1 to 4, wherein the control unit, while immersing the tip of the tip in the liquid of the second container, causes the tip to perform a suction operation so that the multiple cells sucked into the tip move toward the depth of the chip, and then ejects the cells from the tip while moving the head.

10. A cell migration device as described in any one of claims 1 to 4, wherein the control unit causes the tip to perform a suction operation so that the multiple cells sucked into the tip move toward the depth of the tip before the tip of the tip is immersed in the liquid in the second container.

11. A cell migration device as described in claim 3, further comprising a camera capable of capturing an image of the bottom of the first container or the second container, and the control unit having an image processing unit that identifies the containment state of cells in the multiple recesses based on the image captured by the camera.

12. A cell movement device as described in claim 11, wherein the image processing unit identifies an undispersed area at the bottom of the second container that does not contain cells based on the image, and the control unit ejects cells from the tip while moving the head so that the tip passes through the undispersed area.

13. A cell movement device as described in claim 4, further comprising a third container into which target cells are moved from the second container, wherein the control unit identifies a second recess containing a target cell from among the plurality of second recesses, sucks the target cell from the second recess into the tip, moves the head to the third container, and ejects the target cell from the tip into the third container.

14. A cell migration device as described in claim 1, wherein the head is provided in a plurality of units, and the control unit is capable of controlling the operation of the plurality of heads simultaneously or individually.

15. A cell transfer method for transferring a plurality of cells contained in a first container to a second container for storing liquid, comprising the steps of: using a tip having a tip for aspirating and discharging cells, aspirating a plurality of cells into the tip in the first container; moving the tip to the second container; and discharging the cells from the tip while moving the tip with the tip immersed in the liquid in the second container.

16. A cell transfer method as described in claim 15, wherein the first container includes a first bottom that stores liquid and has a plurality of first recesses, and the second container includes a second bottom that has a plurality of second recesses, the tip of the tip, which has previously aspirated a large number of cells, is immersed in the liquid in the first container, and the cells are discharged from the tip while moving the tip, a target recess that contains a target cell to be moved is identified among the plurality of first recesses, all of the cells present in the target recess are aspirated into the tip and the tip is moved to the second container, after the discharging into the second container, the state of cells contained in the plurality of second recesses is confirmed and the second recess in which the target cell is contained is identified, and the target cell is aspirated from the identified second recess by the tip.

Images